Plasmid transformation and replica filter plating of Acholeplasma laidlawii.
The restriction deficient mutant 8195 of Acholeplasma laidlawii strain JA1 was transformed by the promiscuous streptococcal plasmid vector pNZ18 at a frequency of 4 x 10(-4)/cfu. The plasmid was maintained without structural rearrangements but was lost in the absence of a selection pressure, i.e. kanamycin or neomycin. Transformed primary colonies were easily recognized due to a different colony morphology. Replica filter plating, previously not obtained with mycoplasmas, was achieved using pNZ18 as a marker by incubating the replica filters with the cell side down on the new agar plates. These findings should greatly facilitate the genetic and functional analysis of A. laidlawii.
Sundström
T K
TK
Department of Biochemistry, University of Umeå, Sweden.
Wieslander
A
A
D000128:Acholeplasma laidlawii / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D004352:Drug Resistance, Microbial / Q000235:genetics; D007613:Kanamycin Resistance / Q000235:genetics; D009154:Mutation; D009355:Neomycin / Q000494:pharmacology; D010641:Phenotype; D010957:Plasmids; D015183:Restriction Mapping; D014169:Transformation, Bacterial
D009355:Neomycin